Department of Electrical and Computer Engineering, Texas A&M University, College Station, TX 77843, USA.
Lab Chip. 2013 Jun 7;13(11):2144-52. doi: 10.1039/c3lc40877a.
A microfluidic device that simultaneously applies the conditions required for microelectroporation and microsonoporation in a flow-through scheme toward high-efficiency and high-throughput molecular delivery into mammalian cells is presented. This multi-modal poration microdevice using simultaneous application of electric field and ultrasonic wave was realized by a three-dimensional (3D) microelectrode scheme where the electrodes function as both electroporation electrodes and cell flow channel so that acoustic wave can be applied perpendicular to the electric field simultaneously to cells flowing through the microfluidic channel. This 3D microelectrode configuration also allows a uniform electric field to be applied while making the device compatible with fluorescent microscopy. It is hypothesized that the simultaneous application of two different fields (electric field and acoustic wave) in perpendicular directions allows formation of transient pores along two axes of the cell membrane at reduced poration intensities, hence maximizing the delivery efficiency while minimizing cell death. The microfluidic electro-sonoporation system was characterized by delivering small molecules into mammalian cells, and showed average poration efficiency of 95.6% and cell viability of 97.3%. This proof of concept result shows that by combining electroporation and sonoporation together, significant improvement in molecule delivery efficiency could be achieved while maintaining high cell viability compared to electroporation or sonoporation alone. The microfluidic electro-sonoporation device presented here is, to the best of our knowledge, the first multi-modal cell poration device using simultaneous application of electric field and ultrasonic wave. This new multi-modal cell poration strategy and system is expected to have broad applications in delivery of small molecule therapeutics and ultimately in large molecule delivery such as gene transfection applications where high delivery efficiency and high viability are crucial.
本文提出了一种微流控装置，该装置以流动方式同时施加微电穿孔和微声穿孔所需的条件，可实现高效、高通量的分子递送入哺乳动物细胞。这种使用电场和超声波同时施加的多模式穿孔微器件是通过三维（3D）微电极方案实现的，其中电极既作为电穿孔电极，又作为细胞流道，从而可以将超声波垂直于电场施加到流过微流控通道的细胞上。这种 3D 微电极结构还允许施加均匀的电场，同时使该装置与荧光显微镜兼容。我们假设同时在两个垂直方向施加两个不同的场（电场和超声波），可以在细胞膜的两个轴上形成瞬时孔，从而在降低穿孔强度的同时最大限度地提高递药效率，同时最大限度地减少细胞死亡。该微流控电声穿孔系统通过将小分子递送入哺乳动物细胞进行了表征，平均穿孔效率为 95.6%，细胞活力为 97.3%。该概念验证结果表明，通过将电穿孔和声穿孔结合在一起，可以在保持高细胞活力的同时，与单独的电穿孔或声穿孔相比，显著提高分子递药效率。据我们所知，本文提出的这种微流控电声穿孔装置是第一个使用电场和超声波同时施加的多模式细胞穿孔装置。这种新的多模式细胞穿孔策略和系统有望在小分子治疗药物的递送上得到广泛应用，最终在大分子递送上得到应用，如基因转染应用，其中高递药效率和高细胞活力至关重要。
